1 Supplementary Table 1. List of antibodies used in western blotting Antibody name Company ...

42 Pages·2012·0.26 MB·English

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Supplementary Table 1. List of antibodies used in western blotting
Antibody name Company Dilution
Anti-p27 Santa Cruz Biotechnology (sc-527) 1:200
Anti- cyclin D1 Santa Cruz Biotechnology (sc-20044) 1:200
Anti-E-cadherin Santa Cruz Biotechnology (sc-71009) 1:1000
Anti-N-cadherin Santa Cruz Biotechnology (sc-59987) 1:1000
Anti- Tubulin Santa Cruz Biotechnology (sc-23948) 1:2000
Anti-Ezrin Santa Cruz Biotechnology (sc-20773) 1:1000
Anti-Cofilin Santa Cruz Biotechnology (sc-33779) 1:1000
Anti-pospho cofilin (ser3) Santa Cruz Biotechnology (sc-21867-R) 1:1000
Anti-LDH-B Santa Cruz Biotechnology (sc-133731) 1:500
Anti- Catenin Santa Cruz Biotechnology (sc-1496) 1:1000
Anti-pospho-Akt (ser473) Cell Signaling (4051) 1:1000
Anti-phospho-myosin light chain 2 (ser19) Cell Signaling (3671) 1:1000
Anti-pospho-Src (tyr416) Cell Signaling (2101) 1:1000
Anti-pospho-p44/42 MAPK (Erk 1/2)
Cell Signaling (4370) 1:2000
(thr202/tyr204)

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Supplementary Table 2. Primer sequences used in the study.
Primer blast program (http://www.ncbi.nlm.nih.gov/tools/primer-blast/) was used to design specific primer sequences.
Name Sequence Length Tm
Forward primer GTGGGCGATGTGGGCCAGAC
Cofilin 280 60°
Reverse primer CCAGGGTGCAGCGGTCCTTG
Forward primer GTGCGAGATCATCGGGCGGG
LIMK1 143 60°
Reverse primer AACAGCGCACGGTGATGGGG
Forward primer TGACTGGTGGTCGGTTGGGGT
ROCK1 271 60°
Reverse primer TCGGAGCGTTTCCCAAGCCC
Forward primer TGGCCGACGCCATCAACACC
Vimentin 257 60°
Reverse primer CACCTCGACGCGGGCTTTGT
Forward primer ACCACCTCCACAGCCACCGT
E-cadherin 518 60°
Reverse primer GTCCAGTTGGCACTCGCCCC
Forward primer ATTGTGGGTGCGGGGCTTGG
N-cadherin 344 60°
Reverse primer GGGTGTGGGGCTGCAGATCG
Forward primer CCTGCTGGCAGCCATCCCAC
Snail 420 60°
Reverse primer GGCAGCGTGTGGCTTCGGAT
Forward primer ACGCCCAGCTACCCAATGGC
Slug 325 60°
Reverse primer AGGGCGCCCAGGCTCACATA
Forward primer CAGCGCACCCAGTCGCTGAA
Twist 234 60°
Reverse primer CCAGGCCCCCTCCATCCTCC
Forward primer ATGGCCGGGTGGAACGCCTA
Profilin1 413 60°
Reverse primer GAACGCCGAAGGTGGGAGGC
Forward primer CCACGGCTGCTTCCAGCTCC
Actin beta 422 60°
Reverse primer GGAGGGGCCGGACTCGTCAT
Forward primer GCATGGCCTTCCGTGTCCCC
GAPDH 216 60°
Reverse primer CAATGCCAGCCCCAGCGTCA
Forward primer TATTGCCAACCGCAGCCGGG
Keratin 8 534 60°
Reverse primer GCGCCAGAGCCAAAGCTGGA
Forward primer TGGCCATGCGCCAGTCTGTG
Keratin 18 555 60°
Reverse primer GTCCCTCTGCCCGGGTCTGT
Forward primer CGCGGCGTATCCGTGTCCTC
Keratin 19 862 60°
Reverse primer GCGCCTCCGTTTCTGCCAGT
Forward primer GGTGGTCCCGGATCAGCGGT
ANXA1 301 60°
Reverse primer GGCCCTTCATGGCAGCACGA
Forward primer AGCGTGCCCCACCTCCAGAA
ANXA2 195 60°
Reverse primer ATCTCGCGTCCCCTTGCCCT
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Forward primer AGGGCGCGGGAACAAACGAA
ANXA3 445 60°
Reverse primer GCCGGCGTGTTCCTCACACA
Forward primer TGCCCGAGAACCCACCCTCC
Transgelin 328 60°
Reverse primer GCCTGGGAGGCCCCTCTGTT
Forward primer GCAGTGCTGTGCCCTCCCAG
CLIC1 394 60°
Reverse primer CCCGGGCGTAGGCATTGCTC
Forward primer GGCTTGTGGCCGAAGGGCAT
Vinculin 394 60°
Reverse primer GTACCAACCGCAGCCGCCTT
Forward primer AGCAAGAAGGGGAGAGTCGGC
LDH-B 258 60°
Reverse primer GGCAGCTGCTGGGATGAATGC
Forward primer GCCGCTAGAGGTGAAATTCTTG
18S 66 60°
Reverse primer CATTCTTGGCAAATGCTTTCG
Forward primer GCCTGGCAGTCCGTTCCACC
Eplin 530 60°
Reverse primer TGCAGCCAGGACACCCACCT
Forward primer TCCCCCGCCGAGAAGTGGAG
Fascin 464 60°
Reverse primer AGGTCCATACCCTGGCGCGT

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Supplementary Table 3. Protein spots differentially expressed between MCF-7 and MDA-231 cell lines
Nominal Peptide Ion
Spot Protein Matched Instrument
mass Peptides sequence
number name Peptides type
(MW) m/z z

K.GVTFNVTTVDTK.R 641.15 2
K.NSNPALNDNLEK.G 664.72 2
Spot 1 CLIC1 27248 4 ESI-TRAP
K.LAALNPESNTAGLDIFAK.F 923.06 2
K.VLDNYLTSPLPEEVDETSAEDEGVSQR.K 998.53 3

R.VLDELTLAR.T 515.44 2
R.IVLQIDNAR.L 521.43 2
K.DAEAWFTSR.T 541.71 2
R.LEQEIATYR.S 561.94 2
R.MSVEADINGLR.R + Oxidation (M) 610.95 2
K.NHEEEISTLR.G 614.45 2
R.ALEAANGELEVK.I 622.87 2
Spot 2 K1C19 44079 14 ESI-TRAP
R.SQYEVMAEQNR.K 677.99 2
K.AALEDTLAETEAR.F 695.55 2
R.QSSATSSFGGLGGGSVR.F 778.11 2
R.DYSHYYTTIQDLR.D 559.29 3
R.TLQGLEIELQSQLSMK.A + Oxidation (M) 917.84 2
R.SLLEGQEDHYNNLSASK.V 635.99 3
R.GQVGGQVSVEVDSAPGTDLAK.I 1007.71 2

R.AQIFANTVDNAR.I 660.26 2
R.TVQSLEIDLDSMR.N + Oxidation (M) 761.90 2
Spot 3 K1C18 48029 4
R.GGMGSGGLATGIAGGLAGMGGIQNEK.E + 2 Oxidation (M) 765.23 3 ESI-TRAP
R.YALQMEQLNGILLHLESELAQTR.A + Oxidation (M) 896.36 3

Spot 4 GDIR2 23031 2
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K.TLLGDGPVVTDPK.A 656.11 2 ESI-TRAP
K.ELQEMDKDDESLIK.Y + Oxidation (M) 854.84 2

K.LTQDQDVDVK.Y 580.61 2
K.ELVSDANQHVK.S 620.52 2
R.NLCSDDTPMVR.R 654.17 2 ESI-TRAP
Spot 5 2AAA 66039 6
K.DNTIEHLLPLFLAQLK.D 622.23 3
R.QLSQSLLPAIVELAEDAK.W 642.48 3
R.QLSQSLLPAIVELAEDAK.W 963.4 2

R.ALMLQGVDLLADAVAVTMGPK.G + Oxidation (M) 710.44 3
R.TALLDAAGVASLLTTAEVVVTEIPK.E 1241.68 2
Spot 6 CH60 61157 4 ESI-TRAP
R.TALLDAAGVASLLTTAEVVVTEIPK.E 828.41 3
K.LVQDVANNTNEEAGDGTTTATVLAR.S 854.48 3

K.GVFEVK.S 678.42 1
R.QAASSLQQASLK.L 616.37 2
K.DAGQISGLNVLR.V 622.01 2
K.VQQTVQDLFGR.A 646.06 2
R.AQFEGIVTDLIR.R 681.50 2
K.SDIGEVILVGGMTR.M 723.98 2
R.TTPSVVAFTADGER.L 725.96 2
GRP75 / 73680 /
Spot 7 13 / 8 R.EQQIVIQSSGGLSK.D 737.91 2 ESI-TRAP
ANXA6 75873
R.VINEPTAAALAYGLDK.S 823.54 2
K.NAVITVPAYFNDSQR.Q 847.98 2
K.MEEFKDQLPADECNK.L 618.93 3
R.VEAVNMAEGIIHDTETK.M 619.9 3
R.VEAVNMAEGIIHDTETK.M + Oxidation (HW) 625.18 3
/
K.DAISGIGTDEK.C 553.30 2
R.DAFVAIVQSVK.N 589.27 2
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R.EEGGENLDQAR.E 609.34 2
K.SLEDALSSDTSGHFR.R 811.35 2
K.SLHQAIEGDTSGDFLK.A 573.57 3
K.SLHQAIEGDTSGDFLK.A 859.93 2
K.GLGTDEDTIIDIITHR.S 884.94 2
K.GLGTDEDTIIDIITHR.S 590.56 3

R.QFVTPADVVSGNPK.L 729.75 2
Spot 8 PLST 70811 3 R.VYALPEDLVEVKPK.M 799.83 2 ESI-TRAP
K.EGICALGGTSELSSEGTQHSYSEEEK.Y 929.17 3

K.LMIEMDGTENK.S 640.54 2
K.LMIEMDGTENK.S + Oxidation (M) 648.70 2
K.LMIEMDGTENK.S + 2 Oxidation (M) 656.66 2
R.GNPTVEVDLFTSK.G 703.78 2
R.YISPDQLADLYK.S 713.34 2
K.LAQANGWGVMVSHR.S + Oxidation (M) 771.68 2
K.VNQIGSVTESLQACK.L 817.52 2
Spot 9 ENOA 47169 15 R.AAVPSGASTGIYEALELR.D 902.80 2 ESI-TRAP
K.LAMQEFMILPVGAANFR.E 954.45 2
K.LAMQEFMILPVGAANFR.E + Oxidation (M) 962.54 2
K.DATNVGDEGGFAPNILENK.E 981.04 2
K.DATNVGDEGGFAPNILENK.E 654.51 3
R.SGETEDTFIADLVVGLCTGQIK.T 1177.01 2
R.SGETEDTFIADLVVGLCTGQIK.T 785.32 3
K.DYPVVSIEDPFDQDDWGAWQK.F 1255.50 2

R.YALYDATYETK.E 669.49 2
Spot 10 COF1 18719 6 K.LGGSAVISLEGKPL.- 671.06 2 ESI-TRAP
K.KEDLVFIFWAPESAPLK.S 996.03 2
K.EILVGDVGQTVDDPYATFVK.M 1083.97 2
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K.EILVGDVGQTVDDPYATFVK.M 723.02 3
K.NIILEEGKEILVGDVGQTVDDPYATFVK.M 1021.87 3

Spot 11 THIO 12015 1 K.TAFQEALDAAGDK.L 668.71 2 ESI-TRAP

R.SMQDVVEDFK.N 599.79 2
R.SMQDVVEDFK.N + Oxidation (M) 607.46 2
Spot 12 K2C7 51386 4 ESI-TRAP
R.TAAENEFVVLK.K 611.26 2
R.GQLEALQVDGGR.L 621.99 2

R.SLDMDSIIAEVK.A 660.61 2
R.SLDMDSIIAEVK.A + Oxidation (M) 668.68 2
R.ASLEAAIADAEQR.G 672.78 2
Spot 13 K2C8 53671 6 ESI-TRAP
R.LEGLTDEINFLR.Q 710.31 2
K.LEAELGNMQGLVEDFK.N + Oxidation (M) 905.22 2
R.ELQSQISDTSVVLSMDNSR.S + Oxidation (M) 1063.06 2

K.LSELEAALQR.A 1129.79 1
K.YEELQSLAGK.H 1137.6 1
R.SLDMDSIIAEVK.A 660.92 2
R.SLDMDSIIAEVK.A + Oxidation (M) 668.73 2
R.ASLEAAIADAEQR.G 672.7 2
R.ASLEAAIADAEQR.G 672.98 2
Spot 14 K2C8 53671 17 ESI-TRAP
R.TEMENEFVLIK.K 676.78 2
R.TEMENEFVLIK.K + Oxidation (M) 684.67 2
R.LEGLTDEINFLR.Q 710.52 2
K.LEAELGNMQGLVEDFK.N 896.97 2
K.LEAELGNMQGLVEDFK.N + Oxidation (M) 904.99 2
R.SNMDNMFESYINNLR.R 924.49 2
R.SNMDNMFESYINNLR.R 616.76 3
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R.SNMDNMFESYINNLR.R + Oxidation (M) 932.39 2
R.SNMDNMFESYINNLR.R + Oxidation (M) 932.44 2
R.ELQSQISDTSVVLSMDNSR.S 703.18 3
R.ELQSQISDTSVVLSMDNSR.S 1054.94 2

R.SLDMDSIIAEVK.A + Oxidation (M) 668.79 2
R.ASLEAAIADAEQR.G 672.84 2
R.LEGLTDEINFLR.Q 710.46 2
K.LEAELGNMQGLVEDFK.N + Oxidation (M) 905.06 2
Spot 15 K2C8 53671 8 ESI-TRAP
R.SNMDNMFESYINNLR.R + 2 Oxidation (M) 627.26 3
R.SNMDNMFESYINNLR.R + 2 Oxidation (M) 940.65 2
R.ELQSQISDTSVVLSMDNSR.S + Oxidation (M) 1062.86 2
R.ELQSQISDTSVVLSMDNSR.S + Oxidation (M) 709.24 3

K.GIGTDEFTLNR.I 612.06 2
K.SDTSGDYEITLLK.I 721.82 2
Spot 16 ANXA3 36375 5 K.SLGDDISSETSGDFR.K 793.48 2 ESI-TRAP
K.GAGTNEDALIEILTTR.T 837.57 2
R.DYPDFSPSVDAEAIQK.A 891.81 2

K.LSGLPK.H 614.76 1
R.VIGSGCNLDSAR.F 624.68 2
K.MVVESAYEVIK.L 634.18 2
K.MVVESAYEVIK.L + Oxidation (M) 642.32 2
K.IVADKDYSVTANSK.I 755.86 2
Spot 17 LDHB 36900 11 ESI-TRAP
K.SLADELALVDVLEDK.L 815.42 2
K.LIAPVAEEEATVPNNK.I 848.23 2
K.SLADELALVDVLEDKLK.G 624.48 3
K.GEMMDLQHGSLFLQTPK.I + Oxidation (M) 650.04 3
K.GEMMDLQHGSLFLQTPK.I + 2 Oxidation (M) 655.17 3
K.ITVVGVGQVGMACAISILGK.S + Oxidation (M) 663.60 3
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K.IWHHTFYNELR.V 758.33 2
K.SYELPDGQVITIGNER.F 896.06 2
R.VAPEEHPVLLTEAPLNPK.A 652.11 3
K.DLYANTVLSGGTTMYPGIADR.M + Oxidation (M) 1116.01 2
K.DLYANTVLSGGTTMYPGIADR.M + Oxidation (M) 744.55 3
R.TTGIVMDSGDGVTHTVPIYEGYALPHAILR.L + Oxidation (M) 1067.41 3
/ / /
ACTB / 41737 /
K.TPAQFDADELR.A 631.72 2
Spot 18 ANXA1 / 38714 / 6 / 5 / 3 ESI-TRAP
K.GVDEATIIDILTK.R 694.40 2
ACTC 42019
K.GLGTDEDTLIEILASR.T 852.13 2
R.SEDFGVNEDLADSDAR.A 870.34 2
K.GGPGSAVSPYPTFNPSSDVAALHK.A 786.12 3
/ / /
K.IWHHTFYNELR.V 758.33 2
K.ILTERGYSFVTTAER.E 581.44 3
K.SYELPDGQVITIGNER.F 896.06 2

K.DIISDTSGDFR.K 613.78 2
R.TNQELQEINR.V 622.8 2
K.SLYYYIQQDTK.G 711.37 2
K.SYSPYDMLESIR.K + Oxidation (M) 738.36 2
Spot 19 ANXA2 38808 9 K.GVDEVTIVNILTNR.S 772.2 2 ESI-TRAP
K.GLGTDEDSLIEIICSR.T 889.56 2
R.AEDGSVIDYELIDQDAR.D 954.93 2
R.RAEDGSVIDYELIDQDAR.D 1033.06 2
R.RAEDGSVIDYELIDQDAR.D 689.12 3

VIME / 53676 / R.LGDLYEEEMR.E + Oxidation (M) 635.6 2
Spot 20 8 / 7 ESI-TRAP
TBA1B 50804 K.MALDIEIATYR.K + Oxidation (M) 656.1 2
R.EEAENTLQSFR.Q 662.22 2
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R.SLYASSPGGVYATR.S 714.57 2
R.QVQSLTCEVDALK.G 745.87 2
R.KVESLQEEIAFLK.K 767.33 2
R.ISLPLPNFSSLNLR.E 786 2
R.ETNLDSLPLVDTHSK.R 835.03 2
/ / /
R.LSVDYGK.K 781.06 1
K.YMACCLLYR.G + Oxidation (M) 633.1 2
R.LISQIVSSITASLR.F 744.69 2
R.AVFVDLEPTVIDEVR.T 851.52 2
R.IHFPLATYAPVISAEK.A 878.65 2
K.VGINYQPPTVVPGGDLAK.V 913.38 2
K.TIGGGDDSFNTFFSETGAGK.H 1004.42 2

R.WIDNPTVDDR.G 616.23 2
R.VMLVNSMNTVK.E + Oxidation (M) 625.53 2
K.MTGLVDEAIDTK.S + Oxidation (M) 654.55 2
K.QVATALQNLQTK.T 657.65 2
Spot 21 VINC 124292 9 K.AQQVSQGLDVLTAK.V 729.27 2 ESI-TRAP
R.DPSASPGDAGEQAIR.Q 735.81 2
R.GILSGTSDLLLTFDEAEVR.K 679.06 3
R.GILSGTSDLLLTFDEAEVR.K 1018.51 2
K.LLAVAATAPPDAPNREEVFDER.A 794.53 3

R.TIAQDYGVLK.A 554.38 2
K.ATAVMPDGQFK.D + Oxidation (M) 590.86 2
Spot 22 PRDX1 22324 5 R.LVQAFQFTDK.H 598.95 2 ESI-TRAP
R.QITVNDLPVGR.S 606.49 2
K.KQGGLGPMNIPLVSDPK.R + Oxidation (M) 589.92 3

Spot 23 K2C6B 60067 1 ESI-TRAP
K.YEELQITAGR.H 590.60 2
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